Department of Chemistry, Faculty of Agriculture and Forestry, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland.
Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto (U. Porto), Porto; Interdisciplinary Center for Marine and Environmental Research (CIIMAR), University of Porto (U. Porto), Matosinhos, Portugal.
Cryo Letters. 2024 Jan-Feb;45(1):1-15.
The preservation of the nuclear genome's integrity is paramount for the viability and overall health of cells, tissues, and organisms. DNA, being susceptible to damage under physiological conditions and vulnerable to both endogenous and environmental factors, faces constant threats. To assess DNA damage and repair within individual eukaryotic cells, the comet assay presents itself as a versatile, gel electrophoresis-based, relatively simple, and highly sensitive method. Originally designed to monitor DNA damage and repair within populations of mammalian cells, the comet assay has now found applications across diverse domains, including yeast, protozoa, plants, and invertebrates. This technique has proven invaluable in cryopreservation studies, serving as a valuable adjunct for determining suitable cryopreservation protocols. These protocols encompass choices related to cryoprotectants, sample preparation, as well as storage conditions in terms of time and temperature. In the realm of animal cryopreservation research, the comet assay stands as a gold-standard method for assessing DNA integrity. Nevertheless, when applied in plant-oriented investigations, additional efforts are essential due to the distinct nature of plant cells and associated technical challenges. This review elucidates the fundamental principles underlying the comet assay, discusses its current iterations, and delineates its applications in the cryopreservation of both animal and plant specimens. Moreover, we delve into the primary challenges confronting the comet assay's utility as a monitoring tool in the context of plant sample cryopreservation. https://doi.org/10.54680/fr24110110112.
核基因组完整性的保护对于细胞、组织和生物体的生存能力和整体健康至关重要。在生理条件下,DNA 容易受到损伤,并且容易受到内源性和环境因素的影响,因此面临着持续的威胁。为了评估单个真核细胞内的 DNA 损伤和修复,彗星试验作为一种基于凝胶电泳的多功能、相对简单和高度敏感的方法应运而生。彗星试验最初是为了监测哺乳动物细胞群体中的 DNA 损伤和修复而设计的,现在已经在包括酵母、原生动物、植物和无脊椎动物在内的多个领域得到了应用。这项技术在冷冻保存研究中非常有价值,是确定合适的冷冻保存方案的有力辅助手段。这些方案包括选择合适的冷冻保护剂、样品制备以及在时间和温度方面的储存条件。在动物冷冻保存研究中,彗星试验是评估 DNA 完整性的金标准方法。然而,当应用于植物研究时,由于植物细胞的独特性质和相关的技术挑战,需要额外的努力。本综述阐述了彗星试验的基本原理,讨论了它的当前迭代,并描述了它在动物和植物标本冷冻保存中的应用。此外,我们还探讨了彗星试验作为植物样本冷冻保存监测工具的主要挑战。https://doi.org/10.54680/fr24110110112.